[논문]Identification, Characterization, and Pathogenicity of Colletotrichum Species Causing Anthracnose of Peach in Korea

Lee, Dae Min; Hassan, Oliul; Chang, Taehyun

doi:10.1080/12298093.2020.1763116

Identification, Characterization, and Pathogenicity of Colletotrichum Species Causing Anthracnose of Peach in Korea 원문보기

Mycobiology, v.48 no.3, 2020년, pp.210 - 218

Lee, Dae Min (Department of Ecology & Environmental System, College of Ecology & Environmental Sciences, Kyungpook National University) , Hassan, Oliul (Department of Ecology & Environmental System, College of Ecology & Environmental Sciences, Kyungpook National University) , Chang, Taehyun (Department of Ecology & Environmental System, College of Ecology & Environmental Sciences, Kyungpook National University)

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Peach (Prunus persica L.) is one of the major fruit crops in South Korea, along with apple, persimmon, and Asian pears. Peach anthracnose is a continuing threat to growers and is accountable for enormous economic loss. In July 2018, anthracnose of peach appeared at different peach orchards in Gyeongsangbuk-do region, Korea. The typical anthracnose symptoms (brown, circular, and necrotic lesions) were observed on the fruits. Anthracnose of peach was surveyed in different peach orchards of Gyeongsangbuk-do, and 20 fungal isolates from 19 diseased fruits were collected. Multigene phylogenetic analyses coupled with morphological characteristic analysis approaches were used for identifying the fungal species isolated from diseased fruits. This study confirmed three Colletotrichum species. Based on the results, Colletotrichum siamense are reported for the first time as causal agents of peach anthracnose alongside C. fructicola and C. fioriniae, which has been reported previously. Pathogenicity assays were performed for the three isolates representing all the species identified, and Koch's postulates on detached healthy peach fruits were verified. All the identified species were pathogenic on peach fruits as the typical anthracnose symptoms were reproduced. Significant variations in the virulence were observed among fungal species on peach fruit.

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문제 정의

The objective of this study was morphological and molecular characterization of Colletotrichum species associated with peach anthracnose in Korea and investigation of the virulence of identified Colletotrichum species.

제안 방법

Figure 1. A 50% majority rule consensus tree obtained by Bayesian inference analysis based on ITS, TUB2, GAPDH, CHS-1, and CAL sequence alignment. Bayesian posterior probability values ≥ 0.
Figure 3. A 50% majority rule consensus tree obtained by Bayesian inference analysis based on ITS, TUB2, GAPDH, and CHS-1 sequence alignment. Bayesian posterior probability values ! 0.
[8]. In this study, we combined ITS, TUB2, GAPDH, CHS-1, and CAL sequence data for solid identification of isolates from peach to be either C. siamense or C. fructicola. Many scholars suggested that not all eight gene sequences are required to reliably separate Colletotrichum species within C.
Individual gene sequence data from current isolates and data retrieved from GenBank were aligned with each other using the MUSCLE tool in MEGA 6 [21] and edited manually.
The main objective of this study was to identify Colletotrichum species associated with peach anthracnose using morphological and multi-locus phylogenetic analyses and pathogenicity tests to confirm Koch’s postulates.

대상 데이터

were conducted using the combined sequences of ITS, TUB2, GAPDH, CHS-1, and CAL (Figure 1). The final dataset consisted of 37 taxa and 2391 characters (nucleotide positions), including alignment gap (considered as missing data). The fungal species C.

성능/효과

Colletotrichum pseudoacutatum (CBS 43677) was included as the outgroup in this phylogenetic analysis. A total of 4991 tree samples were subjected to BI analysis to construct 50% majority rule consensus tree and estimate posterior probability. There was no basic difference in BI, ML, and MP tree topological structures.
The following loci were amplified and sequenced: internal transcribed spacers (ITS) and a partial sequence of beta-tubulin (TUB2), calmodulin (CAL), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), actin (ACT), chitin synthase 1 (CHS-1), and the Apn2bulin (TUB2), calmodulin, and the partial mating type (ApMat). Identification of isolates into C. gloeosporioides sensu lato (s.l.) was done using six loci (ITS, TUB2, GAPDH, CHS-1, CAL, and ApMat), whereas identification of isolates into C. acutatum s.l. was done using four loci (ITS, TUB2, GAPDH, and CHS-1). To amplify ITS, TUB2, GAPDH, CHS-1, CAL, and ApMat gene, primer pairs ITS1 + ITS4 [16], Bt2a + Bt2b [17], GDF + GDR [18], CHS-79F + CHS-345R [19], CL1C + CL2C [8] and AM-F + AM-R [20] were used, respectively.
In conclusion, this study provides the first morphological, molecular, and pathological characterization of Colletotrichum species in association with peach anthracnose in South Korea. More specifically, this study is also the first report on peach anthracnose caused by C.
Mycelium plugs (5 mm) were transferred to a fresh PDA plate from the actively growing culture. Morphological characteristics of each colony, including color, mycelial growth rate, and conidia and appressorial characteristics, were evaluated after eight days of incubation at 25℃ in the dark. Conidial mass from the culture was mounted in sterilized distilled water or lactic acid.
The isolates recovered from infected peach fruits were clustered in two clades in the C. gloeosporioides s.l. phylogenetic tree: two isolates formed a clade with the ex-type isolates of C. fructicola (ICMP 18581), whereas another two isolates were clustered with the ex-type isolates of C. siamense (ICMP 18642).
The main objective of this study was to identify Colletotrichum species associated with peach anthracnose using morphological and multi-locus phylogenetic analyses and pathogenicity tests to confirm Koch’s postulates. This study revealed three species belonging to the C. gloeosporioides complex (C. siamense and C. fructicola) and the C. acutatum complex (C. fioriniae). Among these, C.

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Identification, Characterization, and Pathogenicity of Colletotrichum Species Causing Anthracnose of Peach in Korea 원문보기

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Identification, Characterization, and Pathogenicity of Colletotrichum Species Causing Anthracnose of Peach in Korea 원문보기

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